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INTRODUCTION: Nineteen percent of 9 years old Irish children are overweight; seven percent are obese. Our aims were: to examine whether differences exist between paternal self-reported and measured height, weight and BMI in a population representative sample; and to explore paternal perceptions of their own weight status. METHODS: Measures of height and weight for fathers and for their children from the National Longitudinal Study of Children Growing Up in Ireland were obtained using validated methods. RESULTS: Three quarters of fathers (6,263 of 8,568 study children) with a mean age of 42 years (SD 5.04) responded. The mean difference between self-reported and measured weight was -1.03kg (SD=4.52), indicating that weight was underestimated. Obese fathers were more likely to have an obese son (9.4% compared to 5.3% for the full cohort) and an obese daughter (12.4% compared to 7.7%). DISCUSSION: These data suggest that there is a strong relationship between fathers' weights and his childrens' weights. A leading factor in the development of childhood obesity is parental obesity. Targeting overweight and obesity in the child should occur simultaneously with tackling overweight and obesity in the parents; in this study, the fathers.
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Índice de Masa Corporal , Peso Corporal , Autoevaluación Diagnóstica , Padre , Obesidad , Adulto , Estatura , Niño , Humanos , Irlanda , Estudios Longitudinales , Masculino , Sobrepeso , Obesidad InfantilRESUMEN
AIMS: Currently there is debate on how best to manage young infants with tongue-tie who have breastfeeding problems. One of the challenges is the subjectivity of the outcome variables used to assess efficacy of tongue-tie division. This structured review documents how the argument has evolved. It proposes how best to assess, inform and manage mothers and their babies who present with tongue-tie related breastfeeding problems. METHODS: Databases were searched for relevant papers including Pubmed, Medline, and the Cochrane Library. Professionals in the field were personally contacted regarding the provision of additional data. Inclusion criteria were: infants less than 3 months old with tongue-tie and/or feeding problems. The exclusion criteria were infants with oral anomalies and neuromuscular disorders. RESULTS: There is wide variation in prevalence rates reported in different series, from 0.02 to 10.7%. The most comprehensive clinical assessment is the Hazelbaker Assessment Tool for lingual frenulum function. The most recently published systematic review of the effect of tongue-tie release on breastfeeding concludes that there were a limited number of studies with quality evidence. There have been 316 infants enrolled in frenotomy RCTs across five studies. No major complications from surgical division were reported. The complications of frenotomy may be minimised with a check list before embarking on the procedure. CONCLUSIONS: Good assessment and selection are important because 50% of breastfeeding babies with ankyloglossia will not encounter any problems. We recommend 2 to 3 weeks as reasonable timing for intervention. Frenotomy appears to improve breastfeeding in infants with tongue-tie, but the placebo effect is difficult to quantify. Complications are rare, but it is important that it is carried out by a trained professional.
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Lactancia Materna , Frenillo Lingual/cirugía , Anomalías de la Boca/cirugía , Anquiloglosia , Femenino , Humanos , Lactante , Frenillo Lingual/anomalías , Madres , Anomalías de la Boca/etiología , Resultado del TratamientoRESUMEN
The use of mannan-oligosaccharides (MOS) as alternatives to antibiotic growth promoters (AGP) has gained in popularity in recent years due to regulatory restrictions of using AGP in food animal production. Benefits of MOS usage include improvement on animal performance, feed efficiency, and gastrointestinal health. The molecular mechanisms of these functions however are not clear. The goal of the current study was to use a transcriptomics approach to investigate the effects of MOS on the intestinal gene expression profile of young broilers and characterize biological gene pathways responsible for the actions of MOS. One hundred and twenty 1-d-old Cobb 500 broiler chicks were randomly divided into 2 groups and were fed either a standard wheat-soybean meal-based (control) diet or the same diet supplemented with 2.2 g/kg of MOS (Bio-Mos, Alltech, Nicholasville, KY) for 3 wk, followed by jejunal gene expression profiling analysis using chicken-specific Affymetrix microarrays. Results indicated that a total of 672 genes were differentially expressed (P < 0.01 and fold change >1.2) in the jejunum by MOS supplementation. Association analysis indicated that differentially expressed genes are involved in diverse biological functions including energy production, cell death, and protein translation. Expression of 77 protein synthesis-related genes was differentially regulated by MOS in the jejunum. Further pathway analysis indicated that 15 genes related to oxidative phosphorylation were upregulated in the jejunum, and expression of genes important in cellular stress response, such as peroxiredoxin 1, superoxide dismutase 1, and thioredoxin, were also increased by MOS. Differential expression of genes associated with cellular immune processes, including lysozyme, lumican, ß 2-microglobin, apolipoprotein A-1, and fibronectin 1, were also observed in MOS-fed broilers. In summary, this study systematically identified biological functions and gene pathways that are important in mediating the biological effects of MOS in broilers.
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Pared Celular/química , Pollos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Yeyuno/metabolismo , Mananos/farmacología , Levaduras/química , Envejecimiento , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Suplementos Dietéticos , Perfilación de la Expresión Génica , Yeyuno/efectos de los fármacos , Mananos/química , Análisis por Matrices de Proteínas/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinariaRESUMEN
Selenium (Se) is an essential component of at least 25 selenoproteins involved in a multitude of physiological functions, including reproduction. However, relatively little is known about the mechanisms by which Se exerts its physiological effects in reproductive tissue. The objective of this study was to compare the effect of long-term inorganic Se (sodium selenite, SS) and organic yeast-derived Se (Sel-Plex(®), SP) supplementations on tissue Se content and gene expression patterns in the oviduct of broiler-breeder hens. Hens were randomly assigned at 6 weeks of age to one of the three treatments: basal semi-purified diet (control), basal diet+0.3 ppm Se as SP or basal diet+0.3 ppm Se as SS. At 49 weeks, oviduct tissue from hens randomly selected from each treatment (n=7) was analyzed for Se content and gene expression profiles using the Affymetrix Chicken genome array. Gene expression data were evaluated using GeneSpring GX 10.0 (Silicon Genetics, Redwood, CA) and Ingenuity Pathways Analysis software (Ingenuity Systems, Redwood City, CA). Oviduct Se concentration was greater with Se supplementation compared with the control (P≤0.05) but did not differ between SS- and SP-supplemented groups. Gene expression analysis revealed that the quantity of gene transcripts associated with energy production and protein translation were greater in the oviduct with SP but not SS supplementation. Targets up-regulated by SP, but not SS, included genes encoding several subunits of the mitochondrial respiratory complexes, ubiquinone production and ribosomal subunits. SS hens showed a decrease in transcripts of genes involved in respiratory complexes, ATP synthesis and protein translation and metabolism in oviduct relative to control hens. In this study, although tissue Se concentrations did not differ between hens fed SS- and SP-supplemented diets, expression patterns of genes involved in energy production and protein synthesis pathways differed between treatments. These variations may partially explain the differences in reproductive performance reported in hens fed different forms of Se.
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Pollos/genética , Regulación de la Expresión Génica/efectos de los fármacos , Oviductos/efectos de los fármacos , Selenio/administración & dosificación , Animales , Pollos/metabolismo , Biología Computacional , Suplementos Dietéticos , Femenino , Perfilación de la Expresión Génica/métodos , Perfilación de la Expresión Génica/veterinaria , Análisis de Secuencia por Matrices de Oligonucleótidos/veterinaria , Oviductos/metabolismo , Oviductos/fisiología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Distribución Aleatoria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinariaRESUMEN
Previous study indicated that inclusion of an algae-based antioxidant as an antioxidative agent [EconomasE, Alltech, Nicholasville, KY; EcoE] significantly reduced the amount of vitamin E (VE) required in broiler diets without compromising performance and meat quality. To assess the mechanisms related to the VE-saving activity of EcoE, as well as other potential functions related to EcoE and VE supplementation, we analyzed gene expression profiles of breast muscle from broilers fed a control diet, the control diet + 50 IU of VE/kg, the control diet + 100 IU of VE/kg, or the control diet + 200 g of EcoE/ton. Evaluation of the serum antioxidant capacity indicated that dietary supplementation of either a high level of VE (50 or 100 IU of VE/kg) or EcoE significantly improved bird antioxidant status. Analysis of gene expression profiles indicated that expression of 542 genes of the breast muscle were altered (P < 0.05, fold change >1.2) by dietary treatments, of which a significant part were commonly regulated by EcoE and VE (especially the control diet + 50 IU of VE/kg). In addition to the process of cellular oxidation, gene ontology analysis indicated the involvement of EcoE and VE on cell morphology, skeletal and muscular system development and function, immune response, and multiple metabolic processes, including lipid, carbohydrate, and drug metabolism. Results of this experiment indicate that the biological roles of high VE, including its activity as an antioxidant, can be greatly mimicked at the transcriptional level by EcoE, and they suggest a relationship of functional redundancy between VE and EcoE in the broiler diets.
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Antioxidantes/farmacología , Pollos , Dieta/veterinaria , Regulación de la Expresión Génica/efectos de los fármacos , Vitamina E/farmacología , Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Antioxidantes/química , Perfilación de la Expresión Génica/veterinaria , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , LevadurasRESUMEN
Histomoniasis (histomonosis, infectious enterohepatitis, or blackhead) is a disease of turkeys on litter or range caused by the protozoan Histomonas meleagridis, a parasite of worms, primarily spread in feces, in Heterakis gallinarum (cecal worm) eggs, or in Eisenia foetida (earthworms). In this trial, Natustat (Alltech, Inc., Nicholasville, KY), a proprietary plant-derived product, was used at 1.925 kg/tonne and compared with nitarsone in male hybrid turkey diets to 42 days of age on histomonad infected litter (day 7) from broiler breeders. Infected nonsupplemented and uninfected nonsupplemented control groups were also included. Natustat and nitarsone significantly improved 28- and 42-day feed conversion ratios and lowered 28- and 35-day cecal and liver lesion scores compared with infected nonsupplemented turkeys. The body weight at 42 days was greater in the Natustat and nitarsone supplemented groups than in the infected nonsupplemented group.
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Arsenicales/administración & dosificación , Arsenicales/uso terapéutico , Vivienda para Animales , Enfermedades de las Aves de Corral/prevención & control , Enfermedades de las Aves de Corral/parasitología , Infecciones Protozoarias en Animales/prevención & control , Pavos/parasitología , Animales , Antiprotozoarios/administración & dosificación , Antiprotozoarios/uso terapéutico , Peso Corporal , Ciego/patología , Suplementos Dietéticos , Masculino , Plantas MedicinalesRESUMEN
The effects of dietary supplementation of Natustat, a proprietary plant derived product (Alltech Inc., KY, USA) and Salinomycin, on performance, feed efficiency and intestinal lesion scores were observed during two Eimeria challenge trials in broiler chickens. In the first trial chickens were challenged with Eimeria sp. via infecting the litter with a known amount of Eimeria oocysts. In the second trial the source of the Eimeria challenge was the litter from the first trial and the same treatment groups were assigned to the same pens as in the initial trial. Birds were placed 55 per pen with seven pens per treatment. Performance parameters were recorded on days 21 and 42 during both trials. Intestinal lesion scores were assessed on days 14 and 21 during Trial 1 and on day 21 during Trial 2. Average weight gain and feed conversion ratios were significantly improved in the Natustat and Salinomycin treatment groups when compared to the non-supplemented infected group. Furthermore, lesion scores were lower on all sampling days in the Natustat and Salinomycin groups when compared to the non-supplemented group. However, only lesions associated with Eimeria tenella were significantly lowered by Natustat and Salinomycin supplementation. Natustat and Salinomycin were equivalent in alleviating the negative performance effects associated with coccidiosis challenge. In summary, Natustat has the potential to be used as a natural alternative to chemotherapeutic drugs for Eimeria control.
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Coccidiosis/veterinaria , Suplementos Dietéticos , Eimeria , Enfermedades de las Aves de Corral/prevención & control , Animales , Pollos , Coccidiosis/patología , Coccidiosis/prevención & control , Coccidiostáticos/uso terapéutico , Eimeria tenella , Intestinos/patología , Masculino , Piranos/uso terapéutico , Aumento de PesoRESUMEN
Methods are described which facilitate quantification of supplemental cellulase, protease and alpha-amylase when added to animal feedingstuffs at normal industrial inclusion levels. The methods entail extraction of the enzymes from the feedingstuffs by agitation in buffer followed by quantification of extract activity using radial diffusion techniques. A linear relationship between the diameter of the zone of hydrolyzed substrate and the log of the enzyme activity applied is observed over a broad activity range. Assay of a feedingstuff supplemented with 1 kg t(-1) cellulase, protease and alpha-amylase yielded net supplemental activity recoveries of 104+/-11.7%, 91.3+/-6.74% and 126+/-29.5%, respectively. A similar assay method did not prove sufficiently sensitive to facilitate detection of xylanase at typical in-feed inclusion levels. The levels of endogenous cellulase, protease and alpha-amylase activity detected in the unsupplemented feedingstuffs were equivalent to 6.4+/-0.47%, 6.6+/-0.82% and 29.0+/-14.1%, respectively, of a 1 kg t(-1) supplement. The methods are technically straightforward and will facilitate determination of enzyme stabilities during processes such as high-temperature pelleting of feedingstuffs, as well as allowing more rigorous quality control related to enzyme-supplemented animal feedingstuffs.
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Alimentación Animal/análisis , Técnicas de Química Analítica/métodos , Suplementos Dietéticos/análisis , Enzimas/análisis , Celulasa/análisis , Difusión , Endo-1,4-beta Xilanasas/análisis , Hidrólisis , Péptido Hidrolasas/análisis , alfa-Amilasas/análisisRESUMEN
A gene encoding alpha-galactosidase activity was isolated by polymerase chain reaction (PCR) from Saccharomyces cerevisiae NCYC686 and separately placed under the control of transcriptional elements regulating alpha-amylase expression in Aspergillus oryzae and glucoamylase expression in A. awamori. Following transformation of both A. oryzae and A. awamori with their respective expression vectors, induction of heterologous alpha-galactosidase from positively selected clones was effected through the addition of soluble starch (10% wt/vol) to the growth medium. Upon induction in A. oryzae, a transcriptional instability resulted in degradation of mRNA encoding heterologous alpha-galactosidase, thus preventing expression of the active enzyme. The use of a gene fusion strategy in A. awamori overcame this instability and resulted in stable expression of S. cerevisiae alpha-galactosidase. Subsequent to initial (shake flask) experiments, a series of scale-up and optimisation studies led to heterologous expression of the recombinant enzyme in batch fermentation at 51 U mg(-1) total extracellular protein. This was higher than previously published works, which reported extracellular levels of heterologous alpha-galactosidase up to 38 U mg(-1) total protein. Analysis of crude extracts of the fermentation medium revealed significant differences between the activity parameters reported previously in the literature for this enzyme and those observed here. The recombinant enzyme exhibited thermostability properties not previously reported for S. cerevisiae alpha-galactosidase, a trait which would make it suitable for use in processes requiring high temperatures.
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Aspergillus oryzae/genética , Aspergillus/genética , Proteínas Recombinantes/biosíntesis , Saccharomyces cerevisiae/enzimología , alfa-Galactosidasa/biosíntesis , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , alfa-Galactosidasa/química , alfa-Galactosidasa/genéticaAsunto(s)
Antiinfecciosos/farmacología , Antioxidantes/farmacología , Medicamentos Herbarios Chinos/farmacología , Animales , Antibacterianos , Aspergillus/efectos de los fármacos , Bacillus subtilis/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Humanos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
Peroxynitrite (ONOO(-)) is a highly reactive oxidant produced by the interaction of the free radicals superoxide (O*-2) and nitric oxide (NO(*)). In a previous study, we found that peroxynitrite is formed in islet beta-cells of nonobese diabetic (NOD) mice. Here, we report that guanidinoethyldisulphide (GED), a selective inhibitor of inducible nitric oxide synthase (iNOS) and scavenger of peroxynitrite prevents diabetes in NOD mice. GED treatment of female NOD mice, starting at age 5 weeks, delayed diabetes onset (from age 12 to 22 weeks) and significantly decreased diabetes incidence at 30 weeks (from 80% to 17%). GED did not prevent pancreatic islet infiltration by leukocytes; however, beta-cells that stained positive for nitrotyrosine (a marker of peroxynitrite) were significantly decreased in islets of GED-treated mice (1+/-1%) compared with vehicle-treated mice (30+/-9%). In addition, GED significantly inhibited nitric oxide and nitrotyrosine formation and decreased destruction of beta-cells in NOD mouse islets incubated in vitro with the combination of proinflammatory cytokines interleukin 1-beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). These findings indicate that both superoxide and nitric oxide radicals contribute to islet beta-cell destruction in autoimmune diabetes via peroxynitrite formation in the beta-cells.
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Diabetes Mellitus Tipo 1/prevención & control , Inhibidores Enzimáticos/farmacología , Depuradores de Radicales Libres/farmacología , Guanidinas/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Oxidantes/metabolismo , Ácido Peroxinitroso/metabolismo , Animales , Células Cultivadas , Diabetes Mellitus Tipo 1/metabolismo , Inhibidores Enzimáticos/administración & dosificación , Femenino , Depuradores de Radicales Libres/administración & dosificación , Guanidinas/administración & dosificación , Islotes Pancreáticos/citología , Islotes Pancreáticos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos NOD , Óxido Nítrico Sintasa de Tipo II , Páncreas/metabolismo , Páncreas/patologíaRESUMEN
Yucca schidigera Roezl ex Ortgies, family Lillaceae, was fractionated with butan-1-ol to yield a butanol extractable fraction (BE; saponin fraction) and a non-butanol fraction (NBE; non-saponin fraction). Four groups of eight male rats were allowed ad libitum access to diets supplemented with water (control) or 200 mg x kg(-1) total Y. schidigera (TOT) or 200 mg x kg(-1) of each of the fractions (NBE or BE). The effects of dietary supplementation with the fractions and their interactions in TOT were analyzed according to the factorial experimental design by two-way analysis of variance. All three supplementation groups displayed significantly reduced serum urea levels (P < 0.05). The TOT and NBE fractions were found to significantly increase serum insulin levels (P < 0.01) in the absence of any fluctuations in serum glucose levels. Urea cycle enzyme activities, namely, arginase (EC 3.5.3.1) and argininosuccinate lyase (EC 4.3.2.1), were significantly decreased (P < 0.05) in vivo, although no effect was observed in vitro. Both fractions displayed effects, indicating that the active constituents are present in both fractions.
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Insulina/sangre , Liliaceae/metabolismo , Urea/sangre , Alimentación Animal , Animales , Suplementos Dietéticos , Liliaceae/química , Masculino , Ratas , SaponinasRESUMEN
AIMS/HYPOTHESIS: A mechanism implicated in pancreatic islet beta-cell destruction in autoimmune diabetes is the binding of the Fas ligand (FasL) on T cells to Fas receptors on beta cells, causing their destruction. Evidence for this mechanism is, however, controversial. The aim of this study was to find whether the Fas ligand contributes to beta-cell death in autoimmune diabetes. METHODS: We transplanted syngeneic islets under the renal capsule in non-obese diabetic (NOD) mice and treated the mice with a neutralizing monoclonal antibody to the Fas ligand. Survival of beta cells in islet grafts and phenotypes of graft-infiltrating cells were investigated. RESULTS: We found 58% (7 of 12) of mice treated with anti-Fas ligand antibody were normoglycaemic at 30 days after islet transplantation compared with none (0 of 9) of the mice treated with control antibody. Immunohistochemical analysis of islet grafts showed that infiltration of leucocytes (CD4+ T cells, CD8+ T cells, macrophages and neutrophils) and apoptosis of beta cells in the grafts was significantly decreased in mice treated with anti-Fas ligand antibody. Expression of proinflammatory cytokines (interleukin 1 alpha, tumour necrosis factor alpha and interferon gamma) was not different in islet grafts of mice treated with anti-Fas ligand and control antibodies. CONCLUSION/INTERPRETATION: These findings indicate that Fas ligand-mediated mechanisms play a major part in promoting leucocytic infiltration of islets and beta-cell destruction in autoimmune diabetes.
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Anticuerpos Monoclonales/farmacología , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/cirugía , Supervivencia de Injerto/inmunología , Trasplante de Islotes Pancreáticos/inmunología , Glicoproteínas de Membrana/fisiología , Animales , Linfocitos B/inmunología , Linfocitos B/patología , Glucemia/metabolismo , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/patología , Diabetes Mellitus Tipo 1/patología , Proteína Ligando Fas , Femenino , Supervivencia de Injerto/efectos de los fármacos , Trasplante de Islotes Pancreáticos/patología , Leucocitos/inmunología , Leucocitos/patología , Macrófagos/inmunología , Macrófagos/patología , Masculino , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos NOD , Ensayo de Capsula Subrrenal , Factores de Tiempo , Trasplante IsogénicoRESUMEN
AIMS/HYPOTHESIS: Testicular Sertoli cells protect allogeneic islet grafts from rejection after transplantation into animals with chemically induced diabetes. The aims of this study were to determine whether Sertoli cells can protect syngeneic islets from autoimmune destruction after transplantation into non-obese diabetic (NOD) mice and, if so, whether protection is due to Sertoli cell expression of Fas ligand (FasL), believed to be the mechanism that protects against allograft rejection. METHODS: We compared the survival of syngeneic islets transplanted under the renal capsule of nonobese diabetic mice, alone and together with purified Sertoli cells prepared from testes of newborn nonobese diabetic mice. Additionally, we examined the composition of the islet and Sertoli cell co-transplants by immunohistochemistry to determine whether islet graft survival correlated with Sertoli cell expression of Fas ligand. RESULTS: Sertoli cell doses of 1, 2 and 4 x 10(6) cells produced a dose-dependent prolongation of median islet graft survival from 11 days (islets alone) to 32 days (islets + 4 x 10(6) Sertoli cells); addition of 8 x 10(6) Sertoli cells to the islet grafts decreased, however, median survival to 8 days. Immunohistochemical analysis of the islet and Sertoli cell co-transplants showed a correlation between Fas ligand expression by Sertoli cells and graft infiltration by neutrophilic leucocytes, leading to islet beta-cell destruction and diabetes recurrence. CONCLUSION/INTERPRETATION: Sertoli cells exert opposing effects on survival of syngeneic islet grafts in nonobese diabetic mice: Fas ligand-dependent neutrophil infiltration and graft destruction, and Fas ligand-independent protection of the graft from autoimmune destruction.
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Diabetes Mellitus Tipo 1/inmunología , Supervivencia de Injerto , Trasplante de Islotes Pancreáticos/inmunología , Células de Sertoli/inmunología , Animales , Animales Recién Nacidos , Glucemia/metabolismo , Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 1/cirugía , Proteína Ligando Fas , Femenino , Rechazo de Injerto , Recuento de Leucocitos , Masculino , Glicoproteínas de Membrana/análisis , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos NOD , Neutrófilos/inmunologíaRESUMEN
To study the relative roles of CD4(+)and CD8(+)T cells and their cytokine products in autoimmune diabetes development, we selectively depleted CD4(+)and CD8(+)T cells in autoimmune diabetes-prone (DP) biobreeding (BB) rats, by administrations of anti-CD2 and anti-CD8 monoclonal antibody (mAb) respectively. We then analysed cytokine mRNA expression, by PCR assay, in mononuclear leukocytes isolated from islets and spleens of control and mAb-treated DP-BB rats. Depletion of CD4(+)T cells (by anti-CD2 mAb) in blood, spleen and islets prevented diabetes development in DP-BB rats, and depletion of CD8(+)T cells (by anti-CD8 mAb) delayed and significantly decreased diabetes incidence. Depletion of either CD4(+)or CD8(+)T cells completely prevented IFN-gamma mRNA upregulation in islets of DP-BB rats above the low level expressed in islets of diabetes-resistant (DR) BB rats. Also, IL-10 mRNA levels in islets of DP-BB rats were significantly decreased by depletion of either CD4(+)or CD8(+)T cells, whereas the effects of the anti-T cell mAb on mRNA levels of other cytokines in islets (IL-2, IL-4, IL-12p40, and TNF-alpha) were discordant. In contrast, both mAb treatments significantly upregulated IL-4 and TNF-alpha mRNA levels in spleens of DP-BB rats. These results demonstrate that islet infiltration by both CD4(+)and CD8(+)T cells is required for IFN-gamma and IL-10 production in islets and beta-cell destruction. Depletion of either CD4(+)or CD8(+)T cells may prevent beta-cell destruction by decreasing IFN-gamma and IL-10 production in islets and increasing IL-4 and TNF-alpha production systemically.
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Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/inmunología , Interferón gamma/genética , Islotes Pancreáticos/inmunología , Animales , Anticuerpos Monoclonales/administración & dosificación , Suero Antilinfocítico/administración & dosificación , Secuencia de Bases , Citocinas/genética , Cartilla de ADN/genética , Diabetes Mellitus Tipo 1/prevención & control , Expresión Génica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas BB , Bazo/inmunologíaAsunto(s)
Glucemia/análisis , Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 1/cirugía , Interferón gamma/biosíntesis , Interleucina-12/biosíntesis , Trasplante de Islotes Pancreáticos , Animales , Islotes Pancreáticos/metabolismo , Ratones , Ratones Endogámicos NOD , Porcinos , Trasplante HeterólogoRESUMEN
BACKGROUND: The recurrent autoimmune response to syngeneic pancreatic islet grafts transplanted into nonobese diabetic (NOD) mice is cell-mediated and relatively resistant to cyclosporine (CsA) therapy. Therefore, we asked whether interleukin (IL)-4 and IL-10, cytokines that inhibit cell-mediated immunity, might improve the therapeutic effect of CsA. METHODS: We compared the survival of syngeneic islet grafts transplanted into diabetic NOD mice treated with IL-4, IL-10, and CsA, administered as single agents and in combinations. Additionally, we measured mRNA levels of type 1 cytokines (interferon-gamma [IFN-gamma], IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), and transforming growth factor-beta (TGF-beta) to determine whether graft rejection or survival might correlate with expression of these cytokines in the grafts. RESULTS: CsA (20 mg/kg/day) significantly prolonged islet graft survival (median: 20 days vs. 10 days for vehicle-treated mice). Neither IL-4 (2.5 microg, twice daily), nor IL-10 (10 microg, twice daily) significantly prolonged islet graft survival. By contrast, combination therapy with CsA and IL-10 significantly prolonged islet graft survival (median: 34 days) compared with vehicle-treated mice (median: 10 days), and combination therapy with CsA and IL-4 significantly prolonged islet graft survival (median: 59 days) compared with both vehicle-treated mice (median: 10 days) and mice treated with CsA alone (median: 20 days). Islet grafts from normoglycemic mice treated with CsA plus IL-10, and with CsA plus IL-4, were surrounded but not infiltrated by mononuclear leukocytes and beta cells were intact, whereas islet grafts from mice treated with vehicle, IL-4, IL-10, and CsA (as single agents) were infiltrated by mononuclear leukocytes and fewer beta cells were detected. Polymerase chain reaction analysis of cytokine mRNA expression in islet grafts at 8-12 days after transplantation revealed that CsA decreased mRNA levels of type 1 cytokines (IFN-gamma and IL-12p40), whereas CsA plus IL-10 did not, and CsA plus IL-4 increased mRNA levels of IFN-gamma, IL-12p40, and TGF-beta. CONCLUSIONS: These results demonstrate that IL-4, and to a lesser extent IL-10, improves the ability of CsA to prevent autoimmune destruction of beta cells in syngeneic islets transplanted into diabetic NOD mice; however, there is no simple correlation between the protective effects of the different treatment regimens (CsA, CsA plus IL-4, and CsA plus IL-10) and mRNA levels of type 1 cytokines (IFN-gamma, IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), or TGF-beta in the islet grafts.
Asunto(s)
Ciclosporina/uso terapéutico , Supervivencia de Injerto/efectos de los fármacos , Inmunosupresores/uso terapéutico , Interleucina-10/uso terapéutico , Interleucina-4/uso terapéutico , Trasplante de Islotes Pancreáticos , Animales , Glucemia/metabolismo , Ciclosporina/administración & dosificación , Citocinas/biosíntesis , Citocinas/genética , Relación Dosis-Respuesta a Droga , Quimioterapia Combinada , Inmunosupresores/administración & dosificación , Ratones , Ratones Endogámicos NOD , ARN Mensajero/metabolismoRESUMEN
Administration of TNF-alpha to autoimmune diabetes-prone nonobese diabetic mice and biobreeding rats inhibits diabetes development; however, the mechanism(s) of diabetes prevention by TNF-alpha has not been established. We used the model of syngeneic islet transplantation into diabetic nonobese diabetic mice to study the effects of TNF-alpha administration on the types of mononuclear cells and cytokines expressed in the islet grafts and on autoimmune diabetes recurrence. Twice daily i.p. injections of TNF-alpha (20 microg/day) from day 1 to day 30 after islet transplantation significantly prolonged islet graft survival; thus, 70% (16 of 23) of mice treated with TNF-alpha were normoglycemic at 30 days after islet transplantation compared with none (0 of 14) of vehicle-treated control mice. Islet grafts and spleens from TNF-alpha-treated mice at 10 days after islet transplantation contained significantly fewer CD4+ and CD8+ T cells, and significantly decreased mRNA levels of type 1 cytokines (IFN-gamma, IL-2, and TNF-beta) than islet grafts and spleens from control mice. Regarding type 2 cytokines, IL-4 mRNA levels were not changed significantly in islet grafts or spleens of TNF-alpha-treated mice, whereas IL-10 mRNA levels were decreased significantly in islet grafts of TNF-alpha-treated mice and not significantly changed in spleens. TGF-beta mRNA levels in islet grafts and spleens were similar in TNF-alpha-treated and control mice. These results suggest that TNF-alpha partially protects beta cells in syngeneic islet grafts from recurrent autoimmune destruction by reducing CD4+ and CD8+ T cells and down-regulating type 1 cytokines, both systemically and locally in the islet graft.
Asunto(s)
Citocinas/biosíntesis , Diabetes Mellitus Tipo 1/inmunología , Regulación hacia Abajo/inmunología , Supervivencia de Injerto/efectos de los fármacos , Trasplante de Islotes Pancreáticos/inmunología , Factor de Necrosis Tumoral alfa/farmacología , Animales , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/patología , Femenino , Refuerzo Inmunológico de Injertos/métodos , Supervivencia de Injerto/genética , Inmunofenotipificación , Islotes Pancreáticos/patología , Subgrupos Linfocitarios/inmunología , Ratones , Ratones Endogámicos NOD , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas BB , Bazo/metabolismo , Células TH1/inmunología , Trasplante IsogénicoRESUMEN
Cytokines produced by islet-infiltrating mononuclear leukocytes may be involved in islet beta-cell destruction and IDDM. To determine which cytokine(s) might be involved in islet beta-cell destruction, we used a reverse transcriptase-polymerase chain reaction assay to compare levels of cytokine mRNA expression in mononuclear leukocytes freshly isolated from islets of four groups of BB rats aged 60-75 days: diabetes-prone (DP) rats, DP rats protected from diabetes by injection of complete Freund's adjuvant (CFA) at age 25 days, acutely diabetic rats, and diabetes-resistant (DR) rats. We found that islet mononuclear leukocyte levels of gamma-interferon (IFN-gamma) mRNA were significantly higher in DP and diabetic rats than in DR rats, whereas CFA-treated DP rats had similar IFN-gamma mRNA levels to DR rats. Also, interleukin (IL)-2 mRNA levels tended to be higher in islet leukocytes from DP and diabetic rats than from DR rats. Tumor necrosis factor-alpha, IL-4, and IL-10 mRNA levels were not significantly different in islet leukocytes from the four groups of rats. These findings suggest that production of T-helper 1 (Th1)-type cytokines, IFN-gamma and IL-2, by islet-infiltrating cells in BB rats is associated with beta-cell destruction and IDDM development.
Asunto(s)
Citocinas/biosíntesis , Diabetes Mellitus Tipo 1/inmunología , Expresión Génica , Islotes Pancreáticos/inmunología , Leucocitos Mononucleares/inmunología , Enfermedades Pancreáticas/inmunología , Animales , Secuencia de Bases , Cartilla de ADN , Diabetes Mellitus Tipo 1/patología , Diabetes Mellitus Tipo 1/prevención & control , Susceptibilidad a Enfermedades , Adyuvante de Freund/farmacología , Interferón gamma/biosíntesis , Interleucina-10/biosíntesis , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Islotes Pancreáticos/patología , Leucocitos Mononucleares/patología , Datos de Secuencia Molecular , Enfermedades Pancreáticas/patología , Reacción en Cadena de la Polimerasa , ARN Mensajero/biosíntesis , Ratas , Ratas Endogámicas BB , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Bronchocentric granulomatosis (BCTG) is a rare disease associated with bronchial asthma and bronchopulmonary aspergillosis. Idiopathic forms are rarely encountered. We report on a case of BCTG showing clinical, radiological, and cytological evidence suggestive of adenocarcinoma of the lung. The patient is a 69-yr-old female, lifetime nonsmoker with multiple sclerosis who was admitted with a history of ascending cholangitis. Admission chest X-ray documented a 1.5-cm nodule in the left upper lobe of the lung. This was confirmed by CT scan. The lesion was slowly growing. Bronchoscopic examination was normal. Bronchial brushings were inconclusive. A transthoracic fine-needle aspiration showed sheets of highly atypical epithelium with occasional small dyshesive clusters. There was an inflammatory background that was believed to represent tumor diathesis. The cytological interpretation was "suspicious for adenocarcinoma." The patient underwent left upper lobectomy. The lung showed multiple peribronchial granulomas with intense peribronchial lymphoid infiltrate extending into the bronchial mucosa, causing cytological atypia and focal ulceration. Special stains for microorganisms were negative. The patient recovered from surgery and shows no signs of infection. We conclude that BCTG and related lesions can give cytological features that are suggestive of malignancy. Cytological material obtained from these lesions should be interpreted with caution.